KR101670321B1 - Manufacturing method of garlic extract of high quantity Cycloalliin - Google Patents
Manufacturing method of garlic extract of high quantity Cycloalliin Download PDFInfo
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Abstract
본 발명은 마늘 추출물을 효소분해하는 단계를 포함하는 사이클로알리인(cycloalliin) 함량이 증가된 마늘추출물 제조방법 또는 마늘추출물의 사이클로알리인(cycloalliin)함량 증가 방법에 관한 것이다.
본 발명에 따르면, 마늘추출물을 효소분해하거나 유산균발효 시키는 경우 마늘추출물 내에 존재하는 사이클로알리인(cycloalliin)의 함량을 증가시킬 수 있다. 특히, 상기 효소분해 단계 전에 마늘추출물을 유산균 발효시키는 경우, 마늘추출물 내에 존재하는 사이클로알리인(cycloalliin)의 함량을 더욱 증가 시킬 수 있다. 또한, 이러한 추출물을 건강기능식품 첨가물로 사용할 경우, 건강기능식품 제조 시 더 효율적일 수 있다. The present invention relates to a method for producing a garlic extract having an increased cycloalliin content or a method for increasing a cycloalliin content of a garlic extract, comprising the step of enzymatically digesting a garlic extract.
According to the present invention, when the garlic extract is enzymatically decomposed or fermented by lactic acid bacteria, the content of cycloalliin present in the garlic extract can be increased. In particular, when the garlic extract is fermented with lactic acid bacteria before the enzymatic decomposition step, the content of cycloallin present in the garlic extract can be further increased. In addition, when these extracts are used as health functional food additives, they can be more effective in manufacturing health functional foods.
Description
본 발명은 사이클로알리인(cycloalliin) 함량이 증가된 마늘추출물 제조방법 또는 마늘추출물의 사이클로알리인(cycloalliin)함량 증가 방법에 관한 것으로, 더욱 구체적으로 마늘추출물을 효소분해 하는 단계를 포함하고 임의로 상기 효소분해 단계 전에 마늘추출물을 유산균 발효시키는 단계를 더 포함하는 사이클로알리인(cycloalliin)함량이 증가된 마늘추출물 제조방법에 관한 것이다.
The present invention relates to a method for producing a garlic extract having increased cycloalliin content or a method for increasing the content of cycloalliin of garlic extract. More particularly, the present invention relates to a method for producing a garlic extract, And further comprising the step of fermenting the garlic extract with lactic acid bacteria prior to the step of decomposition, wherein the cycloalliin content is increased.
마늘은 고대 이집트 시대부터 재배되었고 유럽에서는 지중해 연안에서 주로 재배하다가 그 뒤 인도와 중국을 걸쳐 우리나라에 전파된 것으로 알려져 있다. 마늘은 파, 골파, 양파, 부추, 달래 등과 함께 식물분류학적으로 백합과 식물로 학명은 알리움 사티붐(Allium sativum)이다. 마늘의 속명 Allium은 원래 냄새를 의미하는 라틴어 Olere에서 유래 되었고 식물학자 Linne가 그 시대의 속명의 Allium을 마늘의 학명으로서 사용한 것이 시작이다. 라틴어의 종명(sativum)은 이 식물이 아주 일찍부터 작물화된 것을 나타내는‘재배(Cultivated)’를 의미하고 있다. 마늘을 뜻하는 영어의 garlic은 gar(창)와 lesc(파)가 합성된 것으로 창 모양을 한 파라는 데서 유래 되었고 우리나라에서는 중국의 명칭을 그대로 사용하였지만 명물기략(名物記略, 1870)에는 맛이 매울 랄(辣, 몹시 매움)하므로‘맹랄(猛辣)’이라 하고 이것이‘마랄’이 되고 다시 마늘이 되었다고 한다.Garlic has been cultivated since the ancient Egyptian period, and it has been known that it was cultivated mainly in the Mediterranean coast in Europe, then spread to India through India and China. Garlic is a botanical taxonomic lily plant with algae, chive, onion, leek, and algae, and its scientific name is Allium sativum. The genus Allium is derived from the Latin Olere, which means the original smell, and the botanist Linne began using Allium, the generic name of that time, as the scientific name for garlic. The Latin name for "sativum" means "Cultivated", which indicates that this plant has been cropped very early. The garlic in English, which means garlic, is derived from a pear shaped window with a gar (window) and a lesc (par). The name of the garlic in Korea was used as it is in Korea, but the name " Because it is mad, it is said to be 'vicious,' and it becomes 'maral' and garlic again.
마늘은 비늘줄기로 되어 있는데, 그 속에는 보호엽, 저장엽, 발아엽 원기가 각각 하나씩 들어있다. 대부분 몸에 꽃줄기가 형성되고 그의 분화는 지하엽맥 착생인편의 분화와 동시에 일어난다. 우리가 식용하는 마늘은 마늘잎의 지하부 엽액에 형성된 인편인데 인편의 착생위치는 종류에 따라 다르며 일반적으로 한 마늘구에 대략 10개 내외의 인편을 가진다.Garlic is made up of scaly stem, which contains a protective leaf, a storage leaf, and a sprout leaf. Most of the stem is formed in the body, and its differentiation occurs at the same time as the division of the facial vein, The garlic we eat is the scallops formed in the underground leaf of garlic leaf. The position of scallop depends on the species and generally has about 10 scales in one garlic sphere.
Cycloalliin은 양파, 마늘 등의 Allium 속 백합과 식물내에 존재하는 기능성 황 화합물 중 하나로 cyclic sulfur imino acid이며 중성지질 합성 억제 효과, 혈행 개선 효과 등을 나타내는 물질로 알려져 있다. 마늘 내 대부분의 기능성 황 화합물은 alliin이라는 전구물질로부터 전환이 이루어지는데 이 alliin은 가공에 의한 조직파쇄 시 마늘 내 효소인 alliinase에 의하여 대부분 allicin으로 전환되며 이로 인해 S-allyl-cysteine, S-methyl-cysteine, S-ethyl-cysteine과 같은 기타 기능성 황 화합물로의 전환이 이루어지지 않는다. 시판 중인 숙성 흑마늘의 경우 조직을 파쇄하지 않은 채 장시간 숙성공정에 의해 alliinase 활성이 억제되어 마늘을 대표하는 기능성 황 화합물인 S-allyl-cysteine의 함량이 마늘추출물에 비하여 높으나 S-allyl-cysteine의 안정성이 낮아 시간이 지남에 따라 함량이 지속적으로 감소하게 된다. 그러나 cycloalliin의 경우 마늘조직 파쇄 시에도 alliinase와 반응하지 않으며 고온에서도 안정한 특성을 지니고 있어 마늘 가공 시에 공정상의 제한이 적으며 가공 이후에도 함량이 일정하게 안정적으로 유지되는 물질특성을 지니고 있다.Cycloalliin is one of the functional sulfur compounds in Allium genus such as onion and garlic and is a cyclic sulfur imino acid. It is known as a substance showing inhibitory effect on neutral lipid synthesis and blood circulation improving effect. Most of the functional sulfur compounds in garlic are converted from precursors called alliin, which is converted to allicin by alliinase, an enzyme in garlic during processing, resulting in the formation of S-allyl-cysteine, S-methyl- cysteine, and other functional sulfur compounds such as S-ethyl-cysteine. The content of S-allyl-cysteine, a functional sulfur compound representing garlic, is higher than that of garlic extract, but the stability of S-allyl-cysteine And the content is continuously decreased over time. However, cycloalliin does not react with alliinase even in the case of garlic tissue disintegration and has stable characteristics even at high temperature, so that there are few limitations in the process of garlic processing and the material content is stable even after processing.
현재 마늘의 경우 안정적인 지표물질이 없어 마늘가공식품의 과학적인 접근 및 개발에 어려움이 있으며 특히 건강기능식품으로의 도입이 어려운 실정이다. 지금까지 지표성분인 cycloallin 함량을 증가시킨 마늘 발효물 제조방법에 관한 보고는 없으며 이를 지표물질로 한 마늘가공식품도 개발되어있지 않은 실정이다. Currently, there is no stable indicator substance in garlic, which makes it difficult to develop scientific approach and development of processed garlic products. So far, there has been no report on the production method of garlic fermented by increasing the content of cycloalin as an indicator, and garlic processed foods using the indicator material have not been developed.
이에, 본 발명자들은 상기 종래기술들의 문제점들을 극복하기 위하여 예의 연구노력한 결과, 마늘추출물을 효소분해 하거나 상기 효소분해 단계 전에 마늘추출물을 유산균 발효시키는 경우, 마늘추출물 내에 존재하는 사이클로알리인(cycloalliin)의 함량을 증가시킬 수 있음을 확인하고, 본 발명을 완성하게 되었다.
The present inventors have made extensive efforts to overcome the problems of the prior art. As a result, when the garlic extract is fermented by enzymatic digestion or by digesting the garlic extract with lactic acid bacteria before the enzymatic digestion step, the inventors of the present invention have found that when cycloaliphatic It is possible to increase the content, and the present invention has been completed.
따라서 본 발명의 주된 목적은 마늘추출물을 효소분해 하는 단계를 포함하는 사이클로알리인(cycloalliin) 함량이 증가된 마늘추출물 제조방법을 제공하는 데 있다.Accordingly, it is a main object of the present invention to provide a method for producing garlic extract having an increased cycloalliin content, comprising the step of enzymatically digesting garlic extract.
본 발명의 다른 목적은 상기 사이클로알리인(cycloalliin) 함량이 증가된 마늘추출물 제조방법 또는 마늘추출물의 사이클로알리인(cycloalliin)함량 증가 방법을 이용한 사이클로알리인(cycloalliin) 함량이 증가된 마늘추출물을 함유하는 식품 및 건강기능식품을 제공하는 데 있다.
Another object of the present invention is to provide a method for producing garlic extract having increased cycloalliin content or garlic extract having increased cycloalliin content by a method of increasing the content of cycloalliin of garlic extract Food and health functional foods.
본 발명의 한 양태에 따르면, 본 발명은 마늘추출물을 효소분해 하는 단계를 포함하는 사이클로알리인(cycloalliin) 함량이 증가된 마늘추출물 제조방법을 제공한다. According to one aspect of the present invention, the present invention provides a method for producing a garlic extract having an increased cycloalliin content, comprising enzymatically decomposing the garlic extract.
본 발명의 제조방법에 있어서, 상기 마늘추출물을 효소분해 하는 단계 전에 유산균발효 시키는 단계를 더 포함하는 것을 특징으로 한다. In the method of the present invention, the method further comprises a step of fermenting the lactic acid bacteria before the enzymatic digestion of the garlic extract.
본 발명의 실시예에 따르면, 상기 마늘추출물을 유산균발효 단계 후 셀룰라아제로 효소분해 하는 경우, 셀룰라아제 효소분해 단계만 거친 마늘추출물(실시예 1)보다 cycloalliin의 함량이 1.5배 증가하였으며, 특히 유산균발효 및 효소분해 하지 않은 마늘추출물(비교예 2)보다는 2배 이상 증가하는 것을 확인할 수 있다. 이러한 결과로 볼 때, 마늘추출물을 효소분해 단계 이전에 유산균발효 시키는 경우 cycloalliin의 함량이 더 증가된 마늘추출물을 제조할 수 있음을 알 수 있다(실시예 5 및 도 3). According to the embodiment of the present invention, when the garlic extract was enzymatically degraded with cellulase after fermentation with lactic acid bacteria, the content of cycloalliin was 1.5 times higher than that of the garlic extract (Example 1) It was confirmed that it was more than twice as much as that of the garlic extract without enzymatic degradation (Comparative Example 2). From these results, it can be seen that when garlic extract is fermented with lactic acid bacteria before the enzymatic digestion step, garlic extract having an increased content of cycloalliin can be produced (Example 5 and Fig. 3).
본 발명에 제조방법에 있어서, 상기 효소분해 단계에서 사용되는 효소는 셀룰라아제인 것을 특징으로 한다.In the production method according to the present invention, the enzyme used in the enzyme decomposition step is a cellulase.
본 발명의 실시예에 따르면, 마늘추출물을 각종 효소로 분해하는 경우 효소분해 하지 않은 마늘추출물(비교예 1)에 비해 cycloalliin의 함량이 증가 되었으며, 특히 셀룰라아제 효소로 분해하는 경우, 셀룰라아제 효소분해 하지 않은 마늘추출물(비교예 1)보다 cycloalliin의 ?t량이 1.5배 증가하는 것을 확인할 수 있다. 이러한 결과로 볼 때, 마늘추출물을 셀룰라아제로 효소분해 하는 경우 cycloalliin의 함량이 증가된 마늘추출물을 제조할 수 있음을 알 수 있다(실시예 1 및 표 2). According to the embodiment of the present invention, when the garlic extract was decomposed into various enzymes, the content of cycloalliin was increased as compared with that of the garlic extract without the enzyme decomposition (Comparative Example 1). In particular, when decomposed into cellulase enzymes, It was confirmed that the? T amount of cycloalliin was 1.5 times higher than that of garlic extract (Comparative Example 1). From these results, it can be seen that when the garlic extract is enzymatically digested with a cellulase, the garlic extract with increased cycloalliin content can be prepared (Examples 1 and 2).
본 발명의 제조방법에 있어서, 상기 효소분해 마늘추출물을 주정추출하는 단계를 더 포함하는 것을 특징으로 한다.In the method of the present invention, the method further comprises extracting the enzyme-decomposed garlic extract by alcohol extraction.
본 발명의 실시예에 따르면, 효소분해 한 마늘추출물을 주정추출 할 경우 cycloalliin의 함량이 효소분해만 실시한 마늘추출물보다 1.5배 증가하는 것을 확인할 수 있다. 이러한 결과로 볼 때, 마늘추출물을 효소분해 후 주정추출하는 경우 cycloalliin의 함량이 더 증가된 마늘추출물을 제조할 수 있음을 알 수 있다(실시예 2 및 표 3).According to the embodiment of the present invention, when the enzyme-decomposed garlic extract is subjected to alcohol extraction, the content of cycloalliin is 1.5 times higher than that of the enzyme-decomposed garlic extract alone. From these results, it can be seen that when the garlic extract is subjected to enzymatic digestion and alcohol extraction, the content of cycloalliin is further increased (Example 2 and Table 3).
본 발명의 제조방법에 있어서, 상기 효소분해 마늘추출물은 마늘추출물에 셀룰라아제를 가한 후 40 ~ 50℃에서 15 ~ 25시간 효소분해 하는 것을 특징으로 한다. In the preparation method of the present invention, the enzymatically decomposed garlic extract is characterized in that a cellulase is added to the garlic extract, followed by enzymatic degradation at 40 to 50 ° C for 15 to 25 hours.
본 발명의 실시예에 다르면, 항시 효소분해 마늘추출물은 마늘과 물을 가하여 열수 추출한 마늘추출액에 효소를 처리를 통해 분해하여 cycloalliin의 함량이 증가된 마늘추출물을 제조하였다. 또한, 주정추출 시 효소분해를 통해 얻은 추출물의 총량과 동량의 추정을 첨가하여 추출하여 cycloalliin의 함량이 증가된 마늘추출물을 제조하였다. According to the embodiment of the present invention, garlic extract with hot decomposition of garlic and water was prepared by enzymatic hydrolysis of garlic extract, which was always extracted with hot water, to produce garlic extract having increased cycloallin content. In addition, the total amount and the same amount of extracts obtained by enzymatic digestion were extracted and added to extract garlic extract with increased cycloalliin content.
본 발명의 제조방법에 있어서, 상기 유산균발효 단계에서 사용되는 유산균은 락토바실러스 플란타룸(Lactobacillus plantarum)인 것을 특징으로 한다.In the production method of the present invention, the lactic acid bacterium used in the lactic acid fermentation step is Lactobacillus plantarum.
본 발명의 실시예에 따르면, 상기 유산균은 마늘추출물을 유산균 발효하여 cycloalliin의 함량을 증가시키는 균주를 확인 후 균주동정분석을 의뢰하여 동정하였다. 또한, 락토바실러스 플란타룸(Lactobacillus plantarum)로 유산균발효 할 경우 cycloalliin의 함량이 유산균발효 하지 않은 마늘 추출액(비교예 2)보다 1.5 ~ 2.0배 증가하는 것을 확인할 수 있었다. 이러한 결과로 볼 때, 마늘추출물을 유산균발효시킬 경우 cycloalliin의 함량이 증가된 마늘추출물을 제조할 수 있음을 알 수 있었다(실시예 4). According to an embodiment of the present invention, the lactic acid bacteria were identified by isolating strains which increased the content of cycloalliin by lactic acid bacteria fermentation of garlic extract, and then subjected to identification of strains. When lactobacillus fermentation was performed with Lactobacillus plantarum, the content of cycloalliin was 1.5 to 2.0 times higher than that of lactic acid fermented non-fermented garlic extract (Comparative Example 2). From these results, it was found that when garlic extract is fermented with lactic acid bacteria, garlic extract having increased cycloalliin content can be prepared (Example 4).
본 발명의 제조방법에 있어서, 상기 유산균발효 마늘추출물은 마늘추출물에 락토바실러스 플라타룸(Lactobacillus plantarum) 유산균을 접종하여 1 ~ 2일 배양하여 발효시키는 것을 특징으로 한다.In the preparation method of the present invention, the lactic acid bacteria fermented garlic extract is characterized in that the garlic extract is inoculated with Lactobacillus plantarum lactic acid bacteria and cultured for 1 to 2 days for fermentation.
본 발명의 실시예에 따르면, 상기 유산균 발효 마늘추출물은 발효조에 마늘과 물을 가하여 멸균한 후 유산균배양액을 접종하여 cycloalliin의 함량이 증가된 마늘추출물을 제조하였다. 또한, 상기 제조방법을 통해 얻은 마늘추출물을 셀룰라아제 효소를 처리하여 분해하는 단계를 더 포함하여 cycloalliin의 함량이 더 증가된 마늘추출물을 제조하였다.According to the embodiment of the present invention, the garlic extract of fermented lactic acid bacteria was sterilized by adding garlic and water to the fermentation tank, and then the culture of lactic acid bacteria was inoculated to produce a garlic extract having increased cycloallin content. In addition, a step of decomposing the garlic extract obtained through the above-described method by treating with a cellulase enzyme was further included to prepare a garlic extract having an increased content of cycloalliin.
본 발명의 다른 양태에 따르면, 본 발명은 제 1항 내지 제 7항 중 어느 한 항의 방법에 따라 제조된 사이클로알리인(cycloalliin) 함량이 증가된 마늘추출물을 함유하는 식품 및 건강기능식품을 제공한다.According to another aspect of the present invention, there is provided a food and health functional food comprising garlic extract having an increased cycloalliin content prepared according to the method of any one of
본 발명에 따르면, cycloalliin은 cyclic sulfur imino acid로 황 화합물 중 하나로 중성지질 합성억제 효과, 혈행개선 효과 등을 나타내는 물질이다. 마늘 내 대부분의 기성능 황 화합물은 alliin이라는 전구물질로부터 전환이 이루어지며, 이 전구물질은 가공에 의해 파쇄 시 기능성 황 화합물로 전환이 이루어지지 않는다. 그러나, cycloalliin은 마늘조직 파쇄 시에도 변함없이 유지되고 고온에도 안정한 특성을 지니고 있다. 따라서 본 발명의 제조방법에 따라 제조된 cycloalliin 함량이 증가된 마늘추출물을 함유하는 식품 및 건강기능식품의 경우, 제조공정상 제한이 적으며 cycloalliin을 지표물질로 한 마늘기능식품을 제조할 수 있다. According to the present invention, cycloalliin is a cyclic sulfur imino acid, which is one of sulfur compounds and exhibits an effect of inhibiting neutral lipid synthesis and improving blood circulation. Most of the sulfur-containing compounds in garlic are converted from alliin precursors, which are not converted to functional sulfur compounds by processing. However, cycloalliin remains unchanged even during garlic tissue crushing and has stable characteristics at high temperatures. Therefore, in case of food and health functional food containing garlic extract having increased cycloalliin content according to the production method of the present invention, it is possible to produce garlic functional food using cycloalliin as an index material with fewer limit of manufacturing process.
본 발명에 있어서, 상기 추출물은 캅셀, 정제, 분말, 과립, 액상, 환 중에서 선택된 어느 하나에 포함되는 것을 특징으로 한다. In the present invention, the extract is contained in any one selected from the group consisting of capsules, tablets, powders, granules, liquids and rings.
본 발명의 추출물을 식품 첨가물로 사용할 경우에는 이를 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 추출물 이외에 다른 성분은 제형에 따라 당업자가 적의 하게 선택하여 배합할 수 있으며, 유효 성분은 1종 또는 2종 이상을 혼합하여 사용할 수 있다. 혼합양은 사용 목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있다.
When the extract of the present invention is used as a food additive, it can be added as it is or can be used together with other food or food ingredients, and can be suitably used according to a conventional method. The ingredients other than the extract may be appropriately selected and blended by those skilled in the art according to the formulations, and the active ingredients may be used alone or in combination of two or more. The amount of mixing can be suitably determined according to the intended use (prevention, health or therapeutic treatment).
이하, 본 발명을 보다 구체적으로 설명한다.Hereinafter, the present invention will be described more specifically.
본 발명의 cycloalliin 고함량 마늘추출물 제조방법은 추출탱크에서 교반을 수행하며 마늘을 열수 추출함으로써 마늘조직의 분쇄와 동시에 사용하고자하는 효소가 마늘 내부 조직과 반응하기 쉽게 하고 여기에 효소를 가하여 일정시간 반응시킴으로써 cycloalliin의 함량증가를 유도한다. 또한 효소반응 후에 주정을 첨가하여 추출공정을 수행하여 잔존하는 cycloalliin을 최대한 회수함으로써 cycloalliin 고함량 마늘추출물을 제조한다.The method of producing cycloalliin high-content garlic extract of the present invention is characterized in that stirring is performed in an extraction tank and hot water extraction is performed to extract garlic, and the enzyme to be used simultaneously with the pulverization of garlic tissue is easily reacted with the inner tissue of garlic, To increase the content of cycloalliin. After the enzymatic reaction, the extract is further added with the alcohol to recover the remaining cycloallin to obtain a cycloalliin - rich garlic extract.
본 발명의 효소에 따른 cycloalliin 함량증가 여부는 마늘추출물 효소반응 전/후의 액을 고속액체크로마토그래피로 cycloalliin의 함량을 측정함으로써 cycloalliin의 함량을 증가시킬 수 있는 효소를 선별할 수 있다. 주정추출에 따른 cycloalliin 함량증가 여부도 이와 동일하게 추출 전/후 액을 고속액체크로마토그래피로 cycloalliin의 함량을 측정함으로써 cycloalliin의 함량의 증가여부를 확인할 수 있다.The increase of cycloallin content according to the enzyme of the present invention can be determined by measuring the content of cycloalliin by high performance liquid chromatography before and after the garlic extract enzyme reaction to select an enzyme capable of increasing the content of cycloalliin. Similarly, the increase of cycloallin content by alcohol extraction can be confirmed by measuring the content of cycloalliin by high performance liquid chromatography before and after extraction.
본 발명의 cycloalliin 고함량 유산균발효 마늘추출물의 제조방법에 있어서, 마늘추출물은 식품에 사용가능한 물, 에탄올 등과 같은 친수성 유기용매 또는 이들을 적절히 혼합한 용매로 식품공학적으로 공지된 방법, 예컨대 열수 추출, 유기용매 추출 등의 방법을 사용하여 제조할 수 있으나, 마늘추출물에 유산균을 접종하여 발효 및 효소분해 과정을 수행하여야 하므로 바람직하게는 열수 추출 방법을 이용하는 것이 적당하며 열수추출 이후에 유산균발효를 수행하여 cycloalliin 함량을 증가시키고 발효물에 효소를 첨가하여 반응시킴으로써 cycloalliin의 함량증가를 최대한 유도시킨 cycloalliin 고함량 유산균발효 마늘추출물을 제조하고자 한다. In the method for producing the cycloalliin high-content lactic acid fermented garlic extract of the present invention, the garlic extract may be a hydrophilic organic solvent such as water, ethanol or the like which is usable for food or a solvent in which these are suitably mixed, Solvent extraction, etc. However, since the fermentation and enzymatic degradation process must be performed by inoculating the lactic acid bacteria into the garlic extract, it is preferable to use the hot water extraction method. After the hot water extraction, lactic acid fermentation is performed and cycloalliin The objective of this study was to prepare fermented garlic extract of cycloaliphatic lactic acid bacteria with high content of cycloalliin by increasing the content and adding enzyme to the fermented product.
본 발명의 cycloalliin 고함량 유산균발효 마늘추출물의 제조방법에 있어서, 상기 유산균의 배양 및 효소반응은 유산균 접종 후 적절한 조건에서 수행될 수 있으나, 마늘추출물에 접종되는 통성혐기성 세균에 속하는 유산균의 특성을 고려하여 바람직하게는 유산균 접종 후 35 ~ 37℃에서 공기주입 없이 50 ~ 100 rpm의 조건으로 1 ~ 2일간 배양하는 것이 바람직하다. 효소의 경우 마늘추출물의 pH가 5 ~ 5.5 부근이므로 최적 pH가 이에 적합한 효소를 사용하는 것이 바람직하다.In the method for producing the cycloalliin high-content lactic acid bacteria fermented garlic extract of the present invention, the cultivation and enzyme reaction of the lactic acid bacteria can be carried out under appropriate conditions after inoculation of the lactic acid bacteria, but the characteristics of the lactic acid bacteria belonging to the tuberous anaerobic bacteria inoculated into the garlic extract are considered Preferably at a temperature of 35 to 37 DEG C after inoculation with lactic acid bacteria, at 50 to 100 rpm for 1 to 2 days without air injection. In the case of enzymes, the pH of the garlic extract is in the vicinity of 5 to 5.5, so that it is preferable to use an enzyme suitable for the optimum pH.
본 발명의 cycloalliin 고함유 유산균발효 마늘추출물의 제조에 사용하기 적절한 유산균의 선별방법은 다음과 같다. 충분히 숙성한 김치를 믹서로 분쇄하여 착즙하고, 멸균된 생리식염수로 착즙액을 희석한 용액을 유산균 배양에 최적인 Lactobacilli MRS 한천배지에 도말하여 배양한 후, 형성된 콜로니들을 선별한다. 각각의 형성된 콜로니들을 선별하여 Lactobacilli MRS 한천배지에 계대배양하고 이로부터 각각의 콜로니를 Lactobacilli MRS 액체배지에 배양한다. 액체배지에 배양된 유산균을 마늘추출액에 접종한 후 배양하여 1.0×106 CFU/ml 이상의 생균력을 보이는 특이적인 균주를 선별하고, 선별된 균주의 마늘발효물 내 cycloalliin 함량을 고속 액체 크로마토그래피로 분석하여 유의적으로 cycloalliin 함량을 증가시킬 수 있는 균주를 선별한다.The lactic acid bacteria suitable for use in the production of the cycloalliin-rich lactic acid fermented garlic extract of the present invention are as follows. A sufficiently aged kimchi is ground with a mixer, and the juice is diluted with sterilized physiological saline. The solution is cultured on a Lactobacilli MRS agar medium optimum for culturing lactic acid bacteria, and colonies formed are selected. Each formed colony is selected and subcultured in Lactobacilli MRS agar medium, from which each colony is cultured in Lactobacilli MRS liquid medium. Lactic acid bacteria cultured in a liquid medium were inoculated into garlic extract and cultured to select a specific strain having a viability of 1.0 × 10 6 CFU / ml or more. The cycloalliin content in the fermented garlic of the selected strain was determined by high performance liquid chromatography The strains that can significantly increase the cycloallin content are selected by analysis.
본 발명에서는 상기 과정으로 선별된 마늘 추출물에서 특이적으로 높은 생존력을 보이고 cycloalliin을 유의적으로 증가시키는 유산균을 선별하고 균주동정 절차를 통해 균주명을 확인하였다. In the present invention, the lactic acid bacteria showing a high survival rate in the garlic extracts selected by the above process and significantly increasing the cycloallin were selected and the strain name was confirmed through the strain identification procedure.
본 발명의 상기 방법에 따라 제조된 cycloalliin 고함량 유산균발효마늘추출물은 유산균배양 과정을 통해 제조되므로, 마늘 고유의 약리효과, 지표성분의 효능, 유산균의 정장(整腸) 효과를 동시에 가질 수 있는 장점이 있다.Since the cycloalliin-rich lactic acid fermented garlic extract prepared according to the method of the present invention is produced through the cultivation process of lactic acid bacteria, it is possible to have the effect of having the unique pharmacological effect of garlic, the effect of the indicator ingredient, and the dressing effect of the lactic acid bacteria .
본 발명에 따르면, cycloalliin 고함량 마늘추출물 및 유산균발효마늘추출물을 함유하는 식품조성물을 제공한다. 사용되는 마늘은 현재 모든 식품에 사용가능한 천연물로 알려져 있으며, 본 발명의 마늘발효물의 제조에 이용되는 유산균 또한 식품사용에 제한이 없는 균주이다. 사용된 효소는 식품의약품안전청(KFDA)에 식품첨가물로서 등록된 안전한 효소이며 따라서 모든 형태의 식품의 제조에 사용될 수 있다. 현재 국내에서 마늘의 일일섭취량은 약 19.7g/day이며 cycloalliin 고함량 마늘추출물 및 유산균발효마늘추출물의 1g 당 마늘원재료의 양은 2 ~ 3g 으로써 최대 9g/day까지 섭취하여도 용이하다. cycloalliin 고함량 마늘추출물 및 유산균발효마늘추출물의 식품조성물은, 식용 가능한 식품첨가물을 더 포함할 수도 있으며, 캅셀, 정제, 분말, 과립, 액상, 환 형태로 제제할 수 있다.According to the present invention, there is provided a food composition comprising a cycloalliin high-content garlic extract and a fermented garlic extract of lactic acid bacteria. The garlic used is currently known as a natural product which can be used in all foods. The lactic acid bacterium used in the production of the fermented garlic of the present invention is also a strain which is not restricted in food use. The enzyme used is a safe enzyme registered as a food additive in the Food and Drug Administration (KFDA) and can therefore be used in the manufacture of all types of food. At present, the daily intake of garlic is about 19.7g / day in Korea, and the amount of garlic raw material per 1g of high cycloalliin garlic extract and fermented garlic extract of lactic acid bacteria is 2 ~ 3g, which is easy to ingest up to 9g / day. The food composition of cycloalliin high-content garlic extract and fermented garlic extract of lactic acid bacteria may further include edible food additives, and may be formulated into capsules, tablets, powders, granules, liquids, and rings.
본 발명에 사용된 용어 “Cycloalliin 고함량 마늘추출물”은 마늘을 기질로 하여 셀룰라아제 효소로 분해하고 주정추출 함으로써 cycloalliin을 고농도로 함유하는 마늘추출물을 말한다. “Cycloalliin 고함량 유산균발효마늘추출물”은 마늘을 미생물 영양원으로 하여 Lactibacillus plantarum 유산균 배양을 통해 cycloalliin의 함량증가가 이루어진 유산균 대사산물 및 효소반응에 의해 cycloalliin을 고농도로 함유하는 마늘추출물을 의미한다. 또한 용어 “Cycloalliin 고함량”은 추출물의 고형분 60% 기준일 때 0.4 ~ 0.5%의 cycloalliin 함량을 함유하는 것을 의미한다.
As used herein, the term " Cycloalliin high-content garlic extract " refers to a garlic extract containing cycloalliin at a high concentration by decomposition with garlic as a substrate and extracting with alcohol. "Cycloalliin high content fermented garlic extract" means garlic extract containing cycloalliin at high concentration by lactic acid metabolite and enzyme reaction in which the content of cycloalliin is increased by culturing Lactibacillus plantarum lactic acid bacteria with garlic as a microbial nutrient source. The term " Cycloalliin high content " also means that it contains a content of cycloalliin of 0.4 to 0.5% based on the 60% solids content of the extract.
마늘추출물을 효소분해하거나 유산균발효 시키는 경우 마늘추출물 내에 존재하는 사이클로알리인(cycloalliin)의 함량을 증가시킬 수 있다. 특히, 상기 효소분해 단계 전에 마늘추출물을 유산균 발효시키는 경우, 마늘추출물 내에 존재하는 사이클로알리인(cycloalliin)의 함량을 더욱 증가시킬 수 있다. 또한, 이러한 유용성 성분이 증가된 식품원료를 사용할 경우, 건강기능 향상에 더 효과적이며 건강기능식품 제조에 적합할 수 있다.
When the garlic extract is enzymatically degraded or lactobacillus fermented, the content of cycloalliin present in the garlic extract can be increased. In particular, when the garlic extract is fermented with lactic acid bacteria before the enzymatic decomposition step, the content of cycloallin present in the garlic extract can be further increased. In addition, when such an ingredient of increased availability is used, it is more effective in improving health function and may be suitable for the production of health functional foods.
도 1은 유산균 발효에 따른 cycloalliin 함량을 나타낸 것이다.
도 2는 유산균 간균 3의 동정결과를 나타낸 것이다.
도 3은 제조방법에 따른 cycloallin 함량을 나타낸 것이다.
도 3에서, 실시예 1은 셀룰라아제 효소 분해된 것이고, 실시예 3은 셀룰라아제 효소분해 및 주정추출된 것이고, 실시예 4는 유산균 간균3(Lactobacillus plantarum)으로 발효한 것이고, 실시예 5는 유산균 발효 및 셀룰라아제 효소분해된 것이다.Figure 1 shows the content of cycloalliin following lactic acid fermentation.
Fig. 2 shows the results of identification of Lactococcus bacterium 3.
3 shows the content of cycloallin according to the production method.
3, Example 1 is cellulase-degraded, Example 3 is cellulase degradation and alcohol extraction, Example 4 is fermentation with Lactobacillus plantarum, Example 5 is fermentation with lactic acid bacteria, Cellulase is enzymatically degraded.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하기로 한다. 이들 실시예는 단지 본 발명을 예시하기 위한 것이므로, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지는 않는다.
Hereinafter, the present invention will be described in more detail with reference to Examples. These embodiments are only for illustrating the present invention, and thus the scope of the present invention is not construed as being limited by these embodiments.
비교예Comparative Example 1 : 마늘추출물의 제조 1: Preparation of garlic extract
추출조에 깐 마늘 10kg과 상수 20kg을 가하고 온도 90 ~ 100℃, 교반속도 100 ~ 200rpm, 1시간 추출하였다. 추출액을 온도 40 ~ 50℃, 교반속도 100 ~ 200rpm, 16 ~ 20시간 숙성하였다. 숙성액을 90℃ 30분 가열한 후 여과하고 여과액을 고형분 60%까지 농축하여 마늘추출물을 2.5kg 제조하였다. 이를 본 발명의 효소분해 비교예로서 사용하였다.
10 kg of garlic and 20 kg of constant water were added to the extraction tank, and the mixture was extracted at a temperature of 90 to 100 ° C and a stirring speed of 100 to 200 rpm for 1 hour. The extract was aged at a temperature of 40 to 50 DEG C and a stirring speed of 100 to 200 rpm for 16 to 20 hours. The aging solution was heated at 90 DEG C for 30 minutes and then filtered. The filtrate was concentrated to a solid content of 60% to prepare 2.5 kg of garlic extract. This was used as a comparative example of the enzymatic decomposition of the present invention.
비교예Comparative Example 2 : 마늘추출물의 제조 2: Preparation of garlic extract
발효조에 깐 마늘 10kg과 상수 20kg을 가하고 온도 100 ~ 110℃, 교반속도 100 ~ 200rpm, 3시간 멸균하였다. 추출액을 온도 37℃, 교반속도 50 ~ 100rpm, 24시간 숙성하였다. 숙성 후 온도 90 ~ 100℃에서 1시간 열처리하고 추가로 온도 40 ~ 50℃, 교반속도 100 ~ 200rpm, 16 ~ 20시간 숙성하였다. 숙성액을 90℃ 30분 가열한 후 여과하고 여과액을 고형분 60%까지 농축하여 마늘추출물을 2.5kg 제조하였다. 이를 본 발명의 유산균배양 및 효소분해 비교예로서 사용하였다.
10 kg of garlic and 20 kg of constant water were added to the fermentation tank and sterilized at a temperature of 100 to 110 ° C and a stirring speed of 100 to 200 rpm for 3 hours. The extract was aged at a temperature of 37 ° C and a stirring speed of 50 to 100 rpm for 24 hours. After aging, the mixture was heat-treated at a temperature of 90 to 100 ° C for 1 hour and aged at a temperature of 40 to 50 ° C and a stirring speed of 100 to 200 rpm for 16 to 20 hours. The aging solution was heated at 90 DEG C for 30 minutes and then filtered. The filtrate was concentrated to a solid content of 60% to prepare 2.5 kg of garlic extract. This was used as a comparative example for the cultivation of the lactic acid bacteria of the present invention and for the degradation of enzymes.
실시예Example 1 : 마늘추출물의 1: Garlic extract cycloalliincycloalline 함량을 증가시킬 수 있는 효소 선별 Enzyme selection to increase the content
추출조에 깐 마늘 10kg과 상수 20kg을 가하고 온도 90 ~ 100℃, 교반속도 100 ~ 200rpm, 1시간 추출하여 마늘추출물을 제조하였다. 제조한 마늘추출물을 온도 40 ~ 50℃로 냉각하여 3kg씩 분액하고 당분해효소(알파아밀라아제, 베타아밀라아제, 글루코아밀라아제), 단백질분해효소(프로타아제), 섬유소분해효소(셀룰라아제, 크실라아제), 펙틴분해효소(펙티나아제)를 각 분액에 각각 5g씩 혼합하였다. 효소혼합액을 온도 40 ~ 50℃, 교반속도 100 ~ 200rpm, 16 ~ 20시간 조건으로 반응시켜 효소분해를 수행하고 온도 90℃, 30분간 열처리하여 효소 실활 후 여과하여 여과액을 고형분 60%까지 농축하여 추출물을 제조하였다. 각각의 효소로 분해한 마늘추출물의 cycloalliin 함량을 고속액체크로마토그래피로 분석하여 효소분해하지 않은 마늘추출물(비교예1)의 함량과 비교함으로써 cycloalliin 함량을 최대로 증가시키는 셀룰라아제 효소를 선별하였다.
10 kg of garlic and 20 kg of constant water were added to the extraction tank and extracted with garlic extract at a temperature of 90 to 100 ° C and a stirring speed of 100 to 200 rpm for 1 hour. The resulting garlic extract was cooled to 40 to 50 ° C, and 3 kg of each of the extracts was added to the resulting solution. The resulting solution was mixed with an enzyme (alpha amylase, beta amylase, glucoamylase), protease (protease), cellulolytic enzyme (cellulase, xylanase) , And pectinolytic enzyme (pectinase) were mixed with each 5 g of each fraction. The enzymatic mixture was reacted at a temperature of 40 to 50 ° C and a stirring speed of 100 to 200 rpm for 16 to 20 hours to carry out enzymatic decomposition. The enzyme was heat-treated at 90 ° C for 30 minutes to remove the enzyme and then filtered to concentrate the filtrate to a solid content of 60% The extract was prepared. The content of cycloalliin in the garlic extracts decomposed with each enzyme was analyzed by high performance liquid chromatography and compared with the content of the garlic extract (Comparative Example 1) which was not digested with the enzyme, the cellulase enzyme which maximizes the cycloalliin content was selected.
실시예Example 2 : 마늘효소분해액의 주정추출 2: Extraction of garlic enzyme digestion solution
실시예 1에서 제조한 각각의 효소분해액에 액량대비 동량의 95% 주정을 첨가하여 온도 70 ~ 80℃, 교반속도 100 ~ 200rpm, 1시간 추출하였다. 추출액을 여과하고 여과액을 고형분 60%까지 농축하여 추출물을 제조하였다. cycloalliin 함량을 고속액체크로마토그래피로 분석하여 각각의 효소분해 마늘추출물을 주정추출 한 추출물의 cycloalliin 함량을 상대적으로 비교하였다.
An equal volume of 95% alcohol was added to each of the enzymolytic solutions prepared in Example 1 at a temperature of 70 to 80 ° C and a stirring speed of 100 to 200 rpm for 1 hour. The extract was filtered and the filtrate was concentrated to a solid content of 60% to prepare an extract. The content of cycloalliin was analyzed by high performance liquid chromatography.
실시예Example 3 : 셀룰라아제 효소분해 및 주정추출을 통한 3: through cellulase degradation and alcohol extraction cycloalliincycloalline 고함량High content 마늘추출물 제조 Manufacture of garlic extract
추출조에 깐 마늘 10kg과 상수 20kg을 가하고 온도 90 ~ 100℃, 교반속도 100 ~ 200rpm, 1시간 추출하였다. 마늘추출물에 셀룰라아제 효소분말을 50g가하고 온도 40 ~ 50℃, 교반속도 100 ~ 200rpm, 16 ~ 20시간 효소분해 하였다. 효소분해물에 동량의 주정을 가하여 온도 70 ~ 80℃, 교반속도 100 ~ 200rpm, 1시간 추출하고 여과하였다. 여과액을 고형분 60%까지 농축하여 Cycloalliin 고함량 마늘추출물 3.5kg을 제조하였다.
10 kg of garlic and 20 kg of constant water were added to the extraction tank, and the mixture was extracted at a temperature of 90 to 100 ° C and a stirring speed of 100 to 200 rpm for 1 hour. 50 g of the cellulase powder was added to the garlic extract, and the enzyme was digested at a temperature of 40 to 50 ° C and a stirring speed of 100 to 200 rpm for 16 to 20 hours. The same amount of alcohol was added to the enzyme digest, and the mixture was extracted at a temperature of 70 to 80 ° C and a stirring speed of 100 to 200 rpm for 1 hour and filtered. The filtrate was concentrated to a solid content of 60% and 3.5 kg of a high content of Cycloalliin garlic extract was prepared.
실시예Example 4. 마늘추출물에서 특이적으로 생육 및 4. Growth and specificity in garlic extract cycloalliincycloalline 함량을 증가시키는 유산균의 선별 Selection of lactic acid bacteria to increase the content
충분히 숙성한 김치를 믹서로 분쇄하여 착즙하고, 멸균된 생리식염수로 희석한 용액을 Lactobacilli MRS 한천배지(100g 당 포도당 2g, 트윈(Tween) 80 0.1g, 암모늄 시트레이트(ammonium Citrate) 0.2g, 초산염(acetate) 0.5g, 가수황산마그네슘 0.01g, 무수황산망간 0.005g, 2염기 인산칼륨 0.2 g, 비프 추출물 1 g, 효모 추출물 0.5g, 단백질효소처리 펩톤 1 g, 한천 2g)에 도말하여 37℃, 2일 배양하여, 형성된 콜로니 20개를 확보하였다. 각각의 콜로니를 Lactobacilli MRS 한천배지에 획선 접종하여, 37℃에서 2일 배양한 후 검경하여, 균의 형태를 관찰하고 간균 형태의 콜로니 10종, 구균 형태의 콜로니 5종을 선별하였다. 각각의 콜로니를 Lactobacilli MRS 액체배지에 접종하여 37℃에서 1일 정치 배양하여 종배양액을 제조하였다. 33% 마늘멸균액(마늘과 물 33:67(w/v) 추출액을 온도 100 ~ 110℃, 교반속도 100 ~ 200rpm, 3시간 멸균)에 각각의 유산균 종배양액을 접종한 후 온도 37℃, 교반속도 50 ~ 100rpm, 24시간 배양하여, 생균수가 1.0×106CFU/ml 이상을 보이는 균주 8종을 선별하였다.A sufficiently aged kimchi was ground with a mixer, and the solution was diluted with sterilized physiological saline. The solution was added to a Lactobacilli MRS agar medium (2 g of glucose per 100 g, 0.1 g of Tween 80, 0.2 g of ammonium citrate, 0.5 g of anhydrous magnesium sulfate, 0.005 g of anhydrous manganese sulfate, 0.2 g of dibasic potassium phosphate, 1 g of beef extract, 0.5 g of yeast extract, 1 g of protein enzyme-treated peptone, 2 g of agar) , And cultured for 2 days to obtain 20 colonies formed. Each colony was inoculated on a Lactobacilli MRS agar medium, cultured at 37 ° C for 2 days, and examined by microscopic examination. Ten colonies and five colonies were selected. Each colony was inoculated into a Lactobacilli MRS liquid medium and cultured at 37 ° C for 1 day to prepare a seed culture. Each of the lactic acid bacterial species cultures was inoculated in a 33% garlic sterilized liquid (a mixture of garlic and water 33:67 (w / v) extract at a temperature of 100 to 110 ° C and a stirring rate of 100 to 200 rpm for 3 hours) Eight strains showing viable cell counts of 1.0 × 10 6 CFU / ml or more were selected by culturing at a speed of 50 to 100 rpm for 24 hours.
각 균주 마늘배양액을 온도 90 ~ 100℃에서 1시간 살균하여 여과하고 여과액을 고형분 60%까지 농축하여 농축액의 cycloalliin 함량을 고속액체크로마토그래피로 분석함으로써 배양 전 마늘추출물의 cycloalliin 함량과 비교하였다. 고속액체크로마토그래피 분석결과 특이적으로 cycloalliin의 함량을 증가시키는 균주 1종을 확인하였고 (사)한국종균협회에 균주동정분석을 의뢰하여 생화학적 패턴으로 균체를 동정하는 방법인 API 분석을 통해 동정한 결과 락토바실러스 플란타룸(Lactobacillus plantarum)인 것으로 확인하였다.
The culture broth of each strain was sterilized at 90 ~ 100 ℃ for 1 hour, and the filtrate was concentrated to a solid content of 60%. The content of cycloalliin in the concentrate was analyzed by high performance liquid chromatography and compared with the content of cycloalliin in garlic extract before incubation. As a result of high performance liquid chromatography analysis, we identified one strain which increased the content of cycloalliin specifically. (G) Identified by API analysis, which is a method of identification of bacteria by biochemical pattern, The result was confirmed to be Lactobacillus plantarum.
실시예Example 5 : 유산균발효 및 셀룰라아제 효소에 의한 5: Fermentation by lactic acid bacteria and cellulase enzymes cycloalliincycloalline 고함량High content 마늘추출물의 제조 Manufacture of garlic extract
멸균된 Lactobacilli MRS 액체배지 600g에 Lactobacillus plantarum 균주를 접종하여 온도 37℃, 24시간 정치 배양하여 종배양액을 제조하였다. 발효조에 깐마늘 10kg과 상수 20kg을 가하고 온도 100 ~ 110℃, 교반속도 100 ~ 200rpm, 3 시간 멸균하였다. 멸균된 마늘추출액에 종배양액을 접종하여 온도 37℃, 교반속도 50 ~ 100rpm, 24시간 배양하였다. 배양액을 온도 90 ~ 100℃에서 1시간 살균하고 온도를 40 ~ 50℃까지 냉각 후 셀룰라아제 효소분말을 50g가하고 온도 40 ~ 50℃, 교반속도 100 ~ 200rpm, 24시간 효소분해 하였다. 효소분해액을 90℃ 30분 가열하여 효소를 실활 하고 여과하였다. 여과액을 고형분 60%까지 농축하여 cycloalliin 고함량 마늘추출물 4kg을 제조하였다.
Lactobacillus plantarum strain was inoculated in 600 g of sterilized Lactobacilli MRS liquid medium and cultured at 37 ° C for 24 hours to prepare seed culture. 10 kg of garlic and 20 kg of constant water were added to the fermentation tank and sterilized at a temperature of 100 to 110 ° C and a stirring speed of 100 to 200 rpm for 3 hours. The seed culture was inoculated into the sterilized garlic extract and incubated at a temperature of 37 DEG C and a stirring speed of 50 to 100 rpm for 24 hours. The culture broth was sterilized at 90 to 100 ° C for 1 hour, cooled to 40 to 50 ° C, and 50 g of the cellulase enzyme powder was added. The mixture was subjected to enzymatic digestion at a temperature of 40 to 50 ° C and a stirring rate of 100 to 200 rpm for 24 hours. The enzyme digestion solution was heated at 90 DEG C for 30 minutes to inactivate the enzyme and filter. The filtrate was concentrated to a solid content of 60% to prepare 4 kg of high cycloalliin garlic extract.
실험예Experimental Example 1 : 마늘추출물의 효소 및 주정추출에 의한 1: Enzymatic and alcoholic extraction of garlic extract cycloalliincycloalline 함량증가 Increase in content
비교예 1에서 제시된 마늘추출물을 대조로하여 마늘추출물을 당분해효소(알파아밀라아제, 베타아밀라아제, 글루코아밀라아제), 단백질분해효소(프로타아제), 섬유소분해효소(셀룰라아제, 크실라아제), 펙틴분해효소(펙티나아제)로 각각 분해하여 제조한 실시예 1에 의해 제조된 추출물의 cycloalliin 함량을 고속액체크로마토그래피로 분석하였다. 고속액체크로마토그래피 분석조건은 표 1과 같이 수행하였으며 각 추출물의 cycloalliin 함량결과는 표 2와 같았다.The garlic extract was digested with the enzymes (alpha amylase, beta amylase, glucoamylase), proteolytic enzyme (protease), cellulolytic enzyme (cellulase, xylanase), pectin degradation The content of cycloalliin in the extract prepared in Example 1, which was prepared by digesting with enzyme (pectinase), was analyzed by high performance liquid chromatography. The HPLC analysis conditions were as shown in Table 1. The results of cycloallin content of each extract were as shown in Table 2.
또한, 각 효소분해 추출물에 동량의 주정을 첨가하여 실시예 2와 같이 수행하고 각 추출물의 cycloalliin 함량을 고속액체크로마토그래피로 분석을 수행한 결과는 표 3과 같았다.
Table 3 shows the results of analysis of the cycloalliin content of each extract by high-performance liquid chromatography. The results are shown in Table 3. < tb >< TABLE >
그 결과, 상기 표 2에서 나타나는 바와 같이 마늘추출물에 섬유분해효소인 셀룰라아제와 크실라아제 효소를 사용하여 분해하는 경우 cycloalliin의 함량이 마늘추출물(비교예 1)에 비하여 약 1.5배 증가하는 것을 확인하였다. 특히, 섬유분해효소 중에서도 셀룰라아제로 분해 시, cycloalliin의 함량 증가가 가장 많이 증가하였다.
As a result, as shown in Table 2, it was confirmed that when the garlic extract was decomposed with the enzymes of cellulase and xylanase, the content of cycloalliin was increased about 1.5 times as compared with the garlic extract (Comparative Example 1) . Especially, the increase of cycloalliin content was the greatest in the degradation of cellulase.
효 소 명
Enzyme name
주정추출 전 Cycloalliin
함량(%) [고형분 60%기준]
Cycloalliin before alcohol extraction
Content (%) [based on 60% solids]
주정추출 후 Cycloalliin 함량(%) [고형분 60%기준]
Cycloalliin content after alcohol extraction (%) [based on 60% solids]
그 결과, 상기 표 3에서 나타나는 바와 같이 각각의 효소분해 마늘추출물에 주정추출을 추가적으로 수행하는 경우 cycloalliin 함량은 효소종류에 관계없이 주정추출 전에 비하여 동일하게 1.5배 정도 증가하는 것을 확인하였다. 따라서 본 발명에 있어 마늘추출물을 셀룰라아제 효소로 분해 후 주정추출을 수행함에 따라 cycloalliin 함량을 증가시키는 방법을 확인하고, 실시예 3과 같이 이를 적용하여 cycloalliin 함량 0.4% 이상의 cycloalliin 고함량 마늘추출물을 제조하였다.
As a result, as shown in Table 3, it was confirmed that the cycloallin content was increased by 1.5 times as much as that before the extraction of the alcohol regardless of the kinds of the enzymes when the extract of the enzymes was further added to the extract of the enzymes. Therefore, in the present invention, a method of increasing the content of cycloalliin by decomposing garlic extract with a cellulase enzyme followed by extracting the alcohol, and then applying the same as in Example 3, a cycloalliin-rich garlic extract having a cycloallin content of 0.4% or more was prepared .
실험예Experimental Example 2 : 마늘추출물의 2: Garlic extract cycloalliincycloalline 함량을 증가시킬 수 있는 균주의 선별 Selection of strains that can increase the content
실시예 4와 같이 마늘추출액에 김치로부터 분리한 유산균을 배양하여 생균수를 측정한 결과, 표 4와 같이 유산균 간균 3과 유산균 간균 5가 마늘멸균 배양 시 생균수가 유의적으로 증가한 것을 확인하였다. 각 균주의 마늘배양액을 살균, 여과, 농축하여 추출물을 제조하고 각 추출물의 cycloalliin 함량을 고속액체크로마토그래피로 분석한 결과 도 1에서와 같이 유산균 간균 3의 배양물에서 cycloalliin 함량이 1.5 ~ 2.0배 특이적으로 증가하는 것을 확인할 수 있었다. 따라서 유산균 간균 3을 균주동정 실시한 결과 도 2에서와 같이 락토바실러스 플란타룸(Lactobacillus plantarum) 균주임을 확인하였다. 따라서 본 발명에서는 락토바실러스 플란타룸을 발효균주로서 선정하였고 실험예 1의 결과를 토대로 마늘멸균액을 유산균 발효 후 셀룰라아제 효소로 분해하여 추출물을 제조하고 cycloalliin 함량을 고속액체크로마토그래피로 분석한 결과 도 3에서와 같이 대조군으로 사용한 비교예 2와 비교하여 cycloalliin 함량을 2배 이상 증가시킬 수 있었으며 이를 토대로 실시예 5와 같이 수행하여 cycloalliin 함량 0.4% 이상인 cycloalliin 고함량 유산균발효 마늘추출물을 제조할 수 있었다.
As shown in Table 4, when the lactic acid bacteria isolated from kimchi were cultured in the garlic extract as in Example 4, the number of viable cells was significantly increased in the case of lactic acid bacterium 3 and lactic acid bacterium 5 as shown in Table 4. The extracts were prepared by sterilizing, filtering and concentrating the culture broth of each strain. The cycloalliin content of each extract was analyzed by high performance liquid chromatography. As shown in FIG. 1, the content of cycloalliin in the culture of Lactobacillus acidophilus 3 was 1.5-2.0 times As shown in Fig. As a result, it was confirmed that Lactobacillus plantarum was a strain as shown in FIG. Therefore, in the present invention, Lactobacillus plantarum was selected as a fermentation strain. Based on the results of Experimental Example 1, the extract of lactic acid bacteria was digested with a cellulase enzyme and the content of cycloallin was analyzed by high performance liquid chromatography 3, the cycloalliin content could be increased more than 2 times as compared with Comparative Example 2. As a result, a cycloalliin-rich fermented garlic extract having a cycloalliin content of 0.4% or more was produced in the same manner as in Example 5.
(Lactobacillus plantarum)Lactobacillus Bacillus 3
( Lactobacillus plantarum )
Claims (10)
A step of fermenting the garlic extract with lactic acid bacteria; And a second step of enzymatically digesting the fermented garlic extract, wherein the lactic acid bacterium used in the lactic acid fermentation step is Lactobacillus plantarum, and the enzyme used in the enzymatic digestion step is a cellulase or xylanase Wherein the enzyme is an enzyme-degrading enzyme.
The method according to claim 1, wherein an enzyme is added to the garlic extract in step 2, followed by enzymatic degradation at 40 to 50 ° C for 15 to 25 hours to increase the content of cycloalliin.
The method according to claim 1, wherein the garlic extract is fermented by inoculating Lactobacillus plantarum lactic acid bacteria to the garlic extract in step 1 for 1 to 2 days and then fermenting the garlic extract to increase the content of cycloalliin. Way.
A health functional food containing garlic extract having an increased cycloalliin content produced according to the method of any one of claims 1, 6 or 8.
The health functional food according to claim 9, wherein the extract is contained in any one selected from the group consisting of capsules, tablets, powders, granules, liquids and rings.
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